Use of CCI-779 as an antineoplastic agent

ABSTRACT

This invention provides the use of CCI-779 in the treatment of neoplasms.

This application is a continuation of prior U.S. application Ser. No.10/010,584, filed on Nov. 13, 2001, now abandoned which claims thebenefit of provisional US Patent application No. 60/249,077, filed Nov.15, 2000, now abandoned, the entire disclosure of which is herebyincorporated by reference.

BACKGROUND OF THE INVENTION

This invention relates to the use of rapamycin 42-ester with3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid (CCI-779) as anantineoplastic agent.

Rapamycin is a macrocyclic triene antibiotic produced by Streptomyceshygroscopicus, which was found to have antifungal activity, particularlyagainst Candida albicans, both in vitro and in vivo [C. Vezina et al.,J. Antibiot. 28, 721 (1975); S. N. Sehgal et al., J. Antibiot. 28, 727(1975); H. A. Baker et al., J. Antibiot. 31, 539 (1978); U.S. Pat. No.3,929,992; and U.S. Pat. No. 3,993,749]. Additionally, rapamycin alone(U.S. Pat. No. 4,885,171) or in combination with picibanil (U.S. Pat.4,401,653) has been shown to have antitumor activity.

The immunosuppressive effects of rapamycin have been disclosed in FASEB3, 3411 (1989). Cyclosporin A and FK-506, other macrocyclic molecules,also have been shown to be effective as immunosuppressive agents,therefore useful in preventing transplant rejection [FASEB 3, 3411(1989); FASEB 3, 5256 (1989); R. Y. Calne et al., Lancet 1183 (1978);and U.S. Pat. No. 5,100,899]. R. Martel et al. [Can. J. Physiol.Pharmacol. 55, 48 (1977)] disclosed that rapamycin is effective in theexperimental allergic encephalomyelitis model, a model for multiplesclerosis; in the adjuvant arthritis model, a model for rheumatoidarthritis; and effectively inhibited the formation of IgE-likeantibodies.

Rapamycin is also useful in preventing or treating systemic lupuserythematosus [U.S. Pat. No. 5,078,999], pulmonary inflammation [U.S.Pat. No. 5,080,899], insulin dependent diabetes mellitus [U.S. Pat. No.5,321,009], skin disorders, such as psoriasis [U.S. Pat. No. 5,286,730],bowel disorders [U.S. Pat. No. 5,286,731], smooth muscle cellproliferation and intimal thickening following vascular injury [U.S.Pat. Nos. 5,288,711 and 5,516,781], adult T-cell leukemia/lymphoma[European Patent Application 525,960 A1], ocular inflammation [U.S. Pat.No. 5,387,589], malignant carcinomas [U.S. Pat. No. 5,206,018], cardiacinflammatory disease [U.S. Pat. No. 5,496,832], and anemia [U.S. Pat.No. 5,561,138].

The preparation and use of hydroxyesters of rapamycin, includingCCI-779, are disclosed in U.S. Pat. No. 5,362,718.

DESCRIPTION OF THE INVENTION

This invention provides the use of CCI-779 as an antineoplastic agent,and particularly for neoplasms which are refractory to standard therapy,or for whom standard therapy is not appropriate. In particular CCI-779is useful in the treatment of renal cancer, soft tissue cancer, breastcancer, neuroendocrine tumor of the lung, cervical cancer, uterinecancer, head and neck cancer, glioblastoma, non-small lung cell cancer,prostate cancer, pancreatic cancer, lymphoma, melanoma, small cell lungcancer, ovarian cancer, endometrial cancer, and colon cancer.

As used in accordance with this invention, the term “treatment” meanstreating a mammal having a neoplastic disease by providing said mammalan effective amount of CCI-779 with the purpose of inhibiting growth ofthe neoplasm in such mammal, eradication of the neoplasm, or palliationof the neoplasm.

As used in accordance with this invention, the term “providing,” withrespect to providing CCI-779, means either directly administeringCCI-779, or administering a prodrug, derivative, or analog which willform an effective amount of CCI-779 within the body.

As used in accordance with this invention, the term “refractory neoplam”refers to neoplasms in patients which typically had progressed followingtreatment with standard chemotherapy that was appropriate for that givenneoplasm.

The preparation of CCI-779 is described in U.S. Pat. No. 5,362,718,which is hereby incorporated by reference.

The antineoplastic activity of CCI-779 was confirmed in a preclinical invitro and in vivo standard pharmacological test procedure which measuredthe ability of CCI-779 to treat human renal cell cancer (a rapidlyprogressive disease with very limited treatment options), as well as intwo Phase I human clinical trials. The procedures used and resultsobtained are briefly described below.

Preclinical Test Procedures

In vitro test procedure: Renal tumor lines HTB-44 and CRL-1161 wereobtained from the American Tissue Culture Collection (ATCC), Bethesda,Md. SN12-C line was obtained from Dr. J. Fidler, M.D. Anderson Hospital,Houston, Tex. Cells were plated in MEM (Gibco) supplemented with 2 mMglutamine, 1 mM sodium pyruvate, 5 ml penicillin-streptomycin solution,1 mM non-essential amino acid solution, 10% fetal bovine solution. Cells(5×10³) were plated in 96 well plates with a final volume of 200 ml andincubated for 24 hours at 37° C. Log dilutions of CCI-779 beginning at100 μg/ml were then added to the cultures for 48 hours. Over the last 5hours, cells were pulsed with 1 μci ³H-thymidine (New England Nuclear,6.7 ci/m Mol). Cells were then harvested and the degree of thymidineuptake determined by liquid scintillation spectrometry. The IC₅₀ wasdetermined as the concentration that produced 50% of the maximum uptakeof thymidine in control untreated cells.

In vivo test procedure: Female Balb/c nu/nu mice were obtained fromCharles River Labs, Wilmington, Del., at 6–8 weeks of age. Mice(n=10/group) were injected sc with 5×10⁶ cells resuspended in a 50%solution of Matrigel (BD Biosciences) and tumors allowed to develop.When tumor size reached 100 mg, mice were treated orally with CCI-779 at25 mg/kg. CCI-779 was dosed for 5 consecutive days with repeated 14 daycycles throughout the duration of the experiment. The formulation usedfor CCI-779 was a 50% ethanol, 49% phosal, 1% tween 80 vehicle forresuspending CCI-779, where the stock was resuspended into a 1:10dilution of the vehicle prior to dosing. Tumor growth was evaluatedusing a vernier caliper and volume (l×w×h) was converted to mass usingthe formula: l×w²/2.

Results

Human renal cell tumors were cultured in vitro in the presence orabsence of CCI-779 for 3 days and the effect on growth determined by³H-thymidine incorporation of control versus treated cells. Table 1shows that IC₅₀ (50% growth inhibitory concentration) for all 3 linestested was in the low nM range.

TABLE 1 The effect of CCI-779 on the growth of human renal tumor cellsin vitro Renal Tumor Line CCI-779 IC₅₀ (nM) HTB-44 5.0 CRL-1161 2.0SN12-C 5.5

The effect of CCI-779 in two human renal lines (HTB-44 and CRL-1161)were was evaluated in vivo by engrafting tumor cells on the flanks ofnude mice. Once tumors were established at a size of about 100 mg, micewere treated with CCI-779 or a vehicle control. Treatment with CCI-779at 25 mg/kg resulted in significant inhibition of tumor cell growth inthe mice (Table 2).

TABLE 2 Effect of CCI-779 on the growth of human renal tumor cells innude mouse xenografts Tumor Mass (mg) Drug Days Cell Line Treatment 0 721 35 49 55 HTB-44 Control 288 ± 21 219 ± 18  616 ± 55  1095 ± 44  2033± 24  2412 ± 342   7 CCI 290 ± 15 156 ± 13* 252 ± 48* 453 ± 85*  980 ±155* 1050 ± 183* % T/C 101 71 41 41 48 44 CRL-1161 Control 273 ± 18 355± 36  413 ± 60  480 ± 127 546 ± 170 507 ± 156 CCI 272 ± 14 219 ± 16* 226± 17* 200 ± 21* 229 ± 28* 268 ± 30  % T/C 100 62 60 42 42 53 *p value -<.05 % T/C - Treated/Control × 100Clinical Trial

Two single agent (CCI-779) Phase I clinical trials have been conducted.In the first study, CCI-779 was administered as a 30 minute i.v.infusion daily for 5 days, every two to three weeks. In the secondstudy, CCI-779 was administered as a 30 minute i.v. infusion, onceweekly. Both trials were open label, ascending dose, single-arm,multicenter studies. Patients were allowed to continue treatment as longas the CCI-779 was tolerated and there was no evidence of obviousdisease progression. The following eligibility criteria were used:

Inclusion Criteria

-   -   1. Patients with a histologic diagnosis of advanced cancer        (solid tumors and, in the first study, lymphomas) who are        refractory to standard therapy or for whom standard therapy is        not appropriate.    -   2. Measurable or evaluable disease.    -   3. At least 3 weeks since prior chemotherapy and/or radiation        therapy (6 weeks since nitrosoureas or mitomycin C).    -   4. At least 4 weeks since any other investigational agent.    -   5. Age at least 18 years old.    -   6. Adequate bone marrow, renal, and hepatic function.    -   7. Serum cholesterol≦350 mg/dL and triglycerides≦300 mg/dL.    -   8. ECOG performance status 0–2.    -   9. Life expectancy of at least 3 months.    -   10. Signed, dated, witnessed written informed consent.

A total of 63 patients and 24 patients were enrolled in first and secondstudies, respectively. Dose levels ranged from 0.75–24 mg/m² and 7.5–220mg/m², with the daily×5 every 2 weeks and weekly schedules,respectively.

The following summarizes the results that were obtained:

In patients having renal cancer on the weekly schedule, 1 partialresponse (≧50% reduction in tumor size) and 2 minor responses (≧25% but<50% reduction in tumor size) were observed. In renal cancer patients onthe daily×5 schedule, 1 minor response, 1 unconfirmed minor response,and 1 stable disease (<25% increase to <25% reduction in tumor size)lasting approximately 5 months were observed. In patients having softtissue sarcoma on the daily×5 dosage schedule, 1 possible partialresponse, 2 minor responses, and 1 stable disease lasting approximately5½ months were observed. In patients with breast cancer on the weeklydosage schedule, one partial response was observed. In patients withneuroendocrine tumor of the lung on the weekly dosage schedule, onepartial response was observed. In patients having cervical cancer on thedaily×5 dosage schedule, one minor response was observed. In patientshaving uterine cancer receiving the daily×5 dosage schedule, oneunconfirmed minor response was observed. In patients having head andneck cancer receiving the daily×5 dosage schedule, 1 stable disease forapproximately 8½ months was observed. In patients having non-small celllung cancer receiving the daily×5 dosage schedule, one partial responsewas observed. These results are particularly surprising, consideringthat the patients in these studies had advanced cancers that weregenerally refractory to standard treatment, and also considering thatthese were Phase I clinical trials, in which efficacy is often limited,as the primary objective of a Phase I trial is to determine the safetyand tolerability of the drug being evaluated.

Based on the results of the preclinical and clinical test procedures,CCI-779 is useful in treating neoplasms, in particular, refractoryneoplasms. More particularly, CCI-779 is useful in treating treatment ofrenal carcinoma, soft tissue carcinoma, breast cancer, neuroendocrinetumor of the lung, cervical cancer, uterine cancer, head and neckcancer, glioblastoma, non-small cell lung cancer, prostate cancer,pancreatic cancer, lymphoma, melanoma, small cell lung cancer, ovariancancer, endometrial cancer, and colon cancer.

As typical with chemotherapy, dosage regimens are closely monitored bythe treating physician, based on numerous factors including the severityof the disease, response to the disease, any treatment relatedtoxicities, age, and health of the patient. Based on the resultsobtained with CCI-779, it is projected that initial i.v. infusiondosages will be between about 0.1 and 100 mg/m² when administered on adaily dosage regimen, and between about 1 and 1000 mg/m² whenadministered on a weekly dosage regimen. Other dosage regimens andvariations are foreseeable, and will be determined through physicianguidance. It is preferred that CCI-779 is administered by i.v. infusionor orally, preferably in the form of tablets or capsules. Other routesof administration are also feasible, such-as via implants, parenterally(besides i.v., such as intraperitoneal and subcutaneous injections),rectally, intranasally, vaginally, and transdermally.

Dosage regimens are expected to vary according, to the route ofadministration. For example, dosages for oral administration are oftenup to tenfold greater than for i.v. administration. It is anticipatedthat CCI-779 may be administered as the sole active chemotherapeuticagent, or may be part of a chemotherapeutic regimen containing more thanone antineoplastic agent. The use of concomitant chemotherapeutic agentsoften allows for dosage reduction of each particular agent, therebyincreasing the safety margin of the particular agents.

Oral formulations containing the active compounds of this invention maycomprise any conventionally used oral forms, including tablets,capsules, buccal forms, troches, lozenges and oral liquids, suspensionsor solutions. Capsules may contain mixtures of the active compound(s)with inert fillers and/or diluents such as the pharmaceuticallyacceptable starches (e.g. corn, potato or tapioca starch), sugars,artificial sweetening agents, powdered celluloses, such as, crystallineand microcrystalline celluloses, flours, gelatins, gums, etc. Usefultablet formulations may be made by conventional compression, wetgranulation or dry granulation methods and utilize pharmaceuticallyacceptable diluents, binding agents, lubricants, disintegrants, surfacemodifying agents (including surfactants), suspending or stabilizingagents, including, but not limited to, magnesium stearate, stearic acid,talc, sodium lauryl sulfate, microcrystalline cellulose,carboxymethylcellulose calcium, polyvinylpyrrolidone, gelatin, alginicacid, acacia gum, xanthan gum, sodium citrate, complex silicates,calcium carbonate, glycine, dextrin, sucrose, sorbitol, dicalciumphosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride,talc, dry starches and powdered sugar. Preferred surface modifyingagents include nonionic and anionic surface modifying agents.Representative examples of surface modifying agents include, but are notlimited to, poloxamer 188, benzalkonium chloride, calcium stearate,cetostearl alcohol, cetomacrogol emulsifying wax, sorbitan esters,colloidol silicon dioxide, phosphates, sodium dodecylsulfate, magnesiumaluminum silicate, and triethanolamine. Oral formulations herein mayutilize standard delay or time release formulations to alter theabsorption of the active compound(s). The oral formulation may alsoconsist of administering the active ingredient in water or a fruitjuice, containing appropriate solubilizers or emulsifiers as needed.

In some cases it may be desirable to administer the compounds directlyto the airways in the form of an aerosol.

The compounds of this invention may also be administered parenterally orintraperitoneally. Solutions or suspensions of these active compounds asa free base or pharmacologically acceptable salt can be prepared inwater suitably mixed with a surfactant such as hydroxy-propylcellulose.Dispersions can also be prepared in glycerol, liquid polyethyleneglycols and mixtures thereof in oils. Under ordinary conditions ofstorage and use, these preparation contain a preservative to prevent thegrowth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (e.g., glycerol, propylene glycol and liquidpolyethylene glycol), suitable mixtures thereof, and vegetable oils.

For the purposes of this disclosure, transdermal administrations areunderstood to include all administrations across the surface of the bodyand the inner linings of bodily passages including epithelial andmucosal tissues. Such administrations may be carried out using thepresent compounds, or pharmaceutically acceptable salts thereof, inlotions, creams, foams, patches, suspensions, solutions, andsuppositories (rectal and vaginal).

Transdermal administration may be accomplished through the use of atransdermal patch containing the active compound and a carrier that isinert to the active compound, is non toxic to the skin, and allowsdelivery of the agent for systemic absorption into the blood stream viathe skin. The carrier may take any number of forms such as creams andointments, pastes, gels, and occlusive devices. The creams and ointmentsmay be viscous liquid or semisolid emulsions of either the oil-in-wateror water-in-oil type. Pastes comprised of absorptive powders dispersedin petroleum or hydrophilic petroleum containing the active ingredientmay also be suitable. A variety of occlusive devices may be used torelease the active ingredient into the blood stream such as asemi-permeable membrane covering a reservoir containing the activeingredient with or without a carrier, or a matrix containing the activeingredient. Other occlusive devices are known in the literature.

Suppository formulations may be made from traditional materials,including cocoa butter, with or without the addition of waxes to alterthe suppository's melting point, and glycerin. Water soluble suppositorybases, such as polyethylene glycols of various molecular weights, mayalso be used.

1. A method of treating a refractory neoplasm in a mammal in needthereof, which comprises administering to said mammal an effectiveamount of CCI-779, wherein said mammal has been previously treated withstandard chemotherapy and wherein said refractory neoplasm hasprogressed following treatment with said standard chemotherapy, whereinsaid refractory neoplasm is selected from the group consisting of softtissue carcinoma, breast cancer, neuoroendocrine tumor of the lung,cervical cancer, uterine cancer, head and neck cancer, and non-smallcell lung cancer.
 2. The method according to claim 1, wherein therefractory neoplasm is soft tissue carcinoma.
 3. A method of treatingrefractory breast cancer in a mammal in need thereof, which comprisesproviding to said mammal an effective amount of CCI-779, wherein saidmammal has been previously treated with standard chemotherapy andwherein said refractory breast cancer has progressed following treatmentwith said standard chemotherapy.
 4. The method according to claim 1,wherein the refractory neoplasm is a neuroendocrine tumor of the lung.5. The method according to claim 1, wherein the refractory neoplasm iscervical cancer.
 6. The method according to claim 1, wherein therefractory neoplasm is uterine cancer.
 7. The method according to claim1, wherein the refractory neoplasm is a head and neck cancer.
 8. Themethod according to claim 1, wherein the refractory neoplasm isnon-small cell lung cancer.